Inhibition by salivary peptides, histatins on hydrogen peroxide generation from neutrophils.

1.组胺素的分离纯化：建立了肝素亲和层析-反相高效液相色谱法分离纯化人唾液组胺素的方法.组胺素对中性粒细胞产生H_2O_2的抑制豚鼠和人外周血中性粒细胞在刺激剂的作用下产生H_2O_2，并伴有细胞粘附和扩散。H_2O_2的自发产生依赖于温度和Ca^<2+>，在37 ℃时，在30-60 min内达到最大值。组胺素抑制H_2O_2的产生，并呈剂量依赖性。组<50>蛋白3、5和1的IC_值分别为3、5和32 μ M。组胺抑制素释放组胺活性的必需部分，组胺抑制素的9个残基的序列KRHHGYR（或K）RK是肥大细胞释放组胺的结构要求.组胺素的表征和定量测定：建立了一种简单的酸提取-高效液相色谱法测定唾液中组胺素的方法。唾液中总组胺含量范围为8 ~ 56 μ M，平均28 μ M，且受检者年龄有显著性差异。组胺素分泌的昼夜变化未被识别。结果表明，组胺与唾液中的高分子量成分结合。

1. Isolation and purification of histatins.Heparin-affinity chromatography followed by reversedphase HPLC has been developed for the purification of human salivary histatins.2. Inhibition by histatins on H_2O_2 generation from neutrophils.Neutrophils obtained from guinea pig and human peripheral blood generated H_2O_2 accompanied by cellular adherence and spreading in the assence of stimulants. This spontaneous H_2O_2 production was dependent on temperature and Ca^<2+>, reaching the maximum in 30-60 min at 37ﾟC. Histatins inhibited this H_2O_2 generation dose-dependently. The IC_<50> value was 3, 5 and 32 muM for histatins 3, 5 and 1, respectively.3. The essential moiety of histatins for histamine releasing activity.The sequence of 9 residues of histatins, KRHHGYR(or K)RK was the structural requirements for the release of histamine from mast cells.4. Characterization and quantitative determination of histatins.A simple method for the assay of histatins in saliva by acid extraction followed by HPLC has been developed. Total histatin value was ranged from 8 to 56 muM with a mean value of 28 muM, and it was significant difference in age of donors for saliva. Diurnal changes of histatin secretion were not recognized. It was found that the histatins bound to the high molecular weight compondnts in saliva.